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Updated: Jul 1, 2025
SILAC Based Proteomic Characterization of Exosomes from HIV-1 Infected Cells
Published on: March 3, 2017
Identification of potential novel proteomic markers of Leishmania spp.-derived exosomes
Alonso da Silva Lira Filho1,2, Andrea Lafleur1,2, Marcelo Marcet-Palacios3,4
1Department of Microbiology and Immunology, McGill University, Montréal, QC, Canada.
Introduction:
Extracellular vesicles (EVs) are heterogenous cell-derived membrane-bound structures which can be subdivided into three distinct classes according to distinct morphological characteristics, cellular origins, and functions. Small EVs, or exosomes, can be produced by the protozoan parasite Leishmania through the evolutionarily conserved ESCRT pathway, and act as effectors of virulence and drivers of pathogenesis within mammalian hosts. Techniques for the identification of EVs of non-mammalian origin, however, remain inaccurate in comparison to their well-characterized mammalian counterparts. Thus, we still lack reliable and specific markers for Leishmania-derived exosomes, which poses a significant challenge to the field.
Methods:
Herein, we utilized serial differential ultracentrifugation to separate Leishmania-derived EV populations into three distinct fractions. Nanoparticle tracking analysis and transmission electron microscopy were used to validate their morphological characteristics, and bioinformatic analysis of LC-MS/MS proteomics corroborated cellular origins and function.
Discussion:
Proteomic data indicated potential novel proteic markers of Leishmania-derived exosomes, including proteins involved in endosomal machinery and the ESCRT pathway, as well as the parasitic phosphatase PRL-1. Further investigation is required to determine the specificity and sensitivity of these markers.
Insights
Researchers identified potential protein markers in Leishmania-derived exosomes using proteomics. These findings may aid in developing specific markers for these parasitic extracellular vesicles (EVs).
Area of Science:
- Parasitology
- Cell Biology
- Biochemistry
Background:
- Extracellular vesicles (EVs) are cell-derived structures with diverse origins and functions.
- Leishmania parasites produce exosomes via the ESCRT pathway, contributing to virulence.
- Current methods for identifying non-mammalian EVs, like those from Leishmania, lack accuracy, hindering research.
Purpose of the Study:
- To identify novel proteic markers for Leishmania-derived exosomes.
- To characterize Leishmania EV populations and their origins.
Main Methods:
- Serial differential ultracentrifugation was used to isolate Leishmania EV fractions.
- Nanoparticle tracking analysis and transmission electron microscopy validated EV morphology.
- LC-MS/MS proteomics analyzed EV protein content to infer cellular origins and functions.
Main Results:
- Proteomic analysis revealed potential markers in Leishmania exosomes, including endosomal/ESCRT pathway proteins and PRL-1.
- EV populations were successfully separated and characterized morphologically and functionally.
- The study highlights proteins associated with parasitic exosome biogenesis.
Conclusions:
- Novel proteomic candidates for Leishmania exosome markers were identified.
- Further studies are needed to confirm the specificity and sensitivity of these potential markers.
- Accurate identification of Leishmania exosomes is crucial for understanding parasite-host interactions.
